CN109601980A - A kind of PUFA soft capsule and preparation method thereof containing sialic acid - Google Patents

A kind of PUFA soft capsule and preparation method thereof containing sialic acid Download PDF

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CN109601980A
CN109601980A CN201811358736.0A CN201811358736A CN109601980A CN 109601980 A CN109601980 A CN 109601980A CN 201811358736 A CN201811358736 A CN 201811358736A CN 109601980 A CN109601980 A CN 109601980A
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pufa
oil
sialic acid
parts
soft capsule
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李翔宇
陆姝欢
肖敏
胡雄
余道政
汪志明
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Cabio Biotech Wuhan Co Ltd
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Cabio Biotech Wuhan Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115—Fatty acids or derivatives thereof; Fats or oils
    • A23L33/12—Fatty acids or derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
    • A23D9/02—Other edible oils or fats, e.g. shortenings, cooking oils characterised by the production or working-up
    • A23D9/04—Working-up
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115—Fatty acids or derivatives thereof; Fats or oils
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/15—Vitamins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/30—Encapsulation of particles, e.g. foodstuff additives
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

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Abstract

The invention belongs to nutritional supplement and preparation method thereof fields, and in particular to a kind of PUFA soft capsule and preparation method thereof containing sialic acid.Content in the soft capsule includes: the PUFA oil suspension of evenly dispersed sialic acid；The PUFA oil suspension is sialic acid to be added in PUFA oil to be dispersed up to the content for taking oil suspension different parts measurement sialic acid, and the control of content difference obtains within 5%.Soft capsule granule content uniform level provided by the present invention is high, and stability is high, portable and can supplement a variety of nutrition simultaneously, and antioxidant addition is few or does not add, highly-safe, suitable for pregnant baby crowd.The PUFA oil suspension containing sialic acid used in the present invention is conducive to the nutritional supplementation substance of pregnant baby, and prepares easy, material and be easy to get, and moderate cost, product is portable, is easy to save, and is suitable for promoting the use of.

Description

A kind of PUFA soft capsule and preparation method thereof containing sialic acid
Technical field
The invention belongs to nutritional supplement and preparation method thereof fields, and in particular to a kind of PUFA flexible glue containing sialic acid Capsule and preparation method thereof.
Background technique
What Chinese invention patent CN104041827 was disclosed is a kind of DHA microcapsules and preparation method thereof containing sialic acid, Sialic acid extracts gained from bird's nest, and sialic acid is dissolved in water phase, and then prepares the DHA microcapsules containing sialic acid.
There are DHA microcapsule powder, DHA soft capsule and sialic acid powder on the market, but have no the DHA soft capsule containing sialic acid, This programme can provide a kind of soft capsule of nutritional ingredient diversification.
For pregnant woman, baby can be alleviated because of it by taking the DHA soft capsule containing sialic acid in pregnancy period and the gestational period Itself sialic acid synthetase and DHA are insufficient and dependent on the sialic acid and DHA in external source food.
Summary of the invention
The present invention provides a kind of PUFA soft capsule and preparation method thereof containing sialic acid.
The PUFA soft capsule containing sialic acid, content include: the PUFA oil suspension of evenly dispersed sialic acid.
The PUFA oil suspension is prepared in the following manner:
Sialic acid is added in PUFA oil, is dispersed up to the content for taking oil suspension different parts measurement sialic acid, it is outstanding by oil Liquid upper layer and lower layer's sample detection, content difference is within 5%.
The dispersing mode, which uses, first to be sheared, grinds again；
When shearing, 10-35min is carried out under the shear velocity that condition is 10000rpm-20000rpm, preferably condition is 10-20mins is carried out under the shear velocity of 15000rpm-18000rpm.
Condition when grinding are as follows: circular grinding 10- under revolution 1000r/min-4000r/min, 0.2-0.9mm strainer 50min；It preferably, is 2000r/min-3000r/min, the circular grinding 20-30mins under 0.4-0.6mm strainer in revolution.
It is further preferred that carrying out 20min under the shear velocity that condition is 17000rpm when shearing；And/or when grinding Condition be 2000r/min, strainer 0.4mm, circular grinding 30min.
In the preparation method of PUFA oil suspension of the present invention, it is 98% or more that the sialic acid, which selects sialic acid content, Saliva acid product；The sialic acid can may be from hydrolysate, the large intestine of natural bird's nest by preparing or being commercially available The tunning of bacillus, bacillus subtilis tunning.
A kind of preferred method that sialic acid being prepared using Escherichia coli as fermentation thalli is provided herein:
Using Escherichia coli as fermentation thalli, seed liquor is pressed to the inoculum concentration of 1-2%, is added to containing sterilization fermentation culture medium Fermentor in, the formula of the fermentation medium are as follows: peptone 10-15g/L, yeast extract powder 5-10g/L, sodium chloride 10- 15g/L, pH7.0 ferment under conditions of 37 DEG C, by way of flow feeding, are mended with the rate of 2.5-3g/ (L*h) Glucose is filled, puts tank after the 70-75h that continuously ferments, obtains the fermentation liquid containing sialic acid, fermented feed liquid divides in such a way that film filters Bacteria residue is separated out, the liquid being obtained by filtration is obtained using after sterilizing, decoloration, filtering, concentration, crystallization, separation, spray drying, sieving To sialic acid powder；It is preferred that the Escherichia coli are the Escherichia coli of deposit number CCTCC NO:M 201810.
PUFA oil of the present invention may include this fields such as DHA, ARA, linolenic acid, linoleic acid acceptable activity The grease of ingredient；Further, including DHA oil or at least one of ARA oil.
Wherein, the DHA oil may come from the produced product of algae, fungi, yeast or fish oil refines product etc. and is rich in The grease of DHA, it is preferable that content >=35% of DHA in DHA oil；, further preferably algae oil；The DHA oil can pass through preparation Or commercially available acquisition, it is only necessary to meet above-mentioned DHA content requirement.It will be understood by those skilled in the art that in DHA oil, except DHA at Exceptionally, there are still the acceptable ingredients in part this field.
Preferred preparation method as DHA oil: using schizochytrium as fermentation thalli, seed liquor is added to and is gone out containing steam In the fermentor of fermentation medium after bacterium, fermentation substrate includes glucose, yeast extract, sodium glutamate, magnesium ion and calcium Ion；The ratio 3-5% and dissolved oxygen ratio that the seed liquor and fermentation medium are controlled in fermentation process are less than 10%, 25- Tank, centrifugation or filtering are put after 28 DEG C of fermentation 100-120h, are collected thallus, are added 0.2% cellulase and 0.2% compound protein Enzyme carries out breaking-wall cell, is centrifugally separating to obtain DHA crude oil, obtains 22 using degumming, alkali refining, decoloration, precipitation, deodorization etc. The DHA oil of 40% or more carbon acid content.
Preferably, in parts by weight, the fermentation substrate includes glucose 40-120g/L, yeast extract 4-6g/L, paddy ammonia Sour sodium 20-40g/L, sodium chloride 1-5g/L, potassium dihydrogen phosphate 1-5g/L, magnesium sulfate 5-10g/L, calcium chloride 0.5-1g/L, carbonic acid Hydrogen sodium 0.5-1g/L, sodium sulphate 5-10g/L, ammonium sulfate 5-20g/L, potassium chloride 0.5-1g/L, pH are natural.It selects above-mentioned specific DHA oil, can have the function that collaboration effectively with sialic acid compatibility.
The ARA oil is rich in ARA (eicosatetraenoic acid), to be provided as from the produced product of mortiella Pseudomonas Preferably, it is further preferred that in ARA oil, content >=40% of ARA；The ARA oil can be by preparation or commercially available acquisition, only Above-mentioned ARA content requirement need to be met.It will be understood by those skilled in the art that in addition to ARA ingredient, there are still portions in the ARA oil Divide the acceptable ingredient in this field.
It is further preferred that institute's ARA oil is prepared in the following manner: using Mortierella alpina as fermentation thalli (preferably selection preservation Number Mortierella alpina for being CCTCC NO:M2013419), fermentation medium after seed liquor is added to containing steam sterilizing In fermentor, fermentation substrate includes glucose, yeast extract, sodium glutamate, magnesium ion and calcium ion；Kind is controlled in fermentation process Tank, centrifugation or filtering are put after less than 10%, 28 DEG C fermentation 100h of the ratio 5% and dissolved oxygen ratio of sub- liquid and fermentation medium, Thallus is collected, 0.2% cellulase is added and 0.2% compound protease carries out breaking-wall cell, is centrifugally separating to obtain ARA maos Oil, using degumming, alkali refining, decoloration, precipitation, deodorization etc. up to ARA oil.
Preferably, in parts by weight, the fermentation substrate includes glucose 40-120g/L, yeast extract 4-6g/L, paddy ammonia Sour sodium 20-40g/L, sodium chloride 1-5g/L, potassium dihydrogen phosphate 1-5g/L, magnesium sulfate 5-10g/L, calcium chloride 0.5-1g/L, carbonic acid Hydrogen sodium 0.5-1g/L, sodium sulphate 5-10g/L, ammonium sulfate 5-20g/L, potassium chloride 0.5-1g/L, pH are natural.
Preferably, seed culture fluid is inoculated into the fermentation flask equipped with fermentation medium after fermented and cultured, further include by Grade expands culture；
The seed culture medium expanded in incubation step by step are as follows: carbon source 20-50g/l；Nitrogen source 10-20g/l；pH 5.5-8.5, and bacterium is dense reach 15-30% (volumetric concentration) when carry out in next step expand culture；Every grade of incubation time is 24- 50h, cultivation temperature are 25-30 DEG C, and ventilatory capacity is required air per minute in 1-2vvm (L/L.min) i.e. every liter of fermentation liquid Intake is 1-2L, and tank presses 0.05-0.15MPa, and mixing speed is that 150-300 turns/min；Then the seed of above-mentioned acquisition is expanded Big culture solution, which is inoculated into final fermentor, carries out fermented and cultured, inoculum concentration 10-20% (volume ratio), fermentation jar temperature 25-30 DEG C, 200-300 revs/min of mixing speed, in every liter of fermentation liquid of ventilatory capacity 1-2vvm (L/L.min) i.e. per minute required for it is empty Gas intake is 1-2L, and tank presses 0.05-0.15Mpa, cultivates 150-180h, and control during the fermentation by stream plus carbon source Fermentation medium of the carbon source concentration in 5-20g/L, the fermentor in fermentation liquid are as follows: carbon source 30-50g/l；Nitrogen source 10-20g/ l；pH5.5-8.5.Degumming, alkali refining, decoloration, precipitation, deodorization etc. obtain 40% or more Eicosatetraenoic acid content again after broken wall The grease of ARA.
Preferably, also can further include can receive containing this fields such as linolenic acid, linoleic acid in the PUFA oil Active constituent grease；Other PUFA oil such as linolenic acid, linoleic acid may be from this fields such as linseed oil, sunflower oil can Other PUFA oil received.
Preferably, in PUFA oil, the content ratio of DHA oil and ARA oil is (2-3): 1, and DHA oil and oily total of ARA Content accounts at least the 80% of the PUFA oil.
In the preparation method of PUFA oil suspension of the present invention, it is preferable that when shearing, condition is the shearing of 17000rpm 20mins is carried out under speed.
In the preparation method of PUFA oil suspension of the present invention, it is preferable that condition when grinding is 2000r/min, filter Net is 0.4mm, circular grinding 30mins.
In the preparation method of PUFA oil suspension of the present invention, it is preferable that the mass ratio of the sialic acid and PUFA oil For 1:(1.5-10), preferably 1: (4-8).
In the preparation method of PUFA oil suspension of the present invention, obtained oil suspension partial size is≤34 μm, it is further excellent Selection of land, the partial size of oil suspension are≤25.5 μm.
PUFA soft capsule of the present invention containing sialic acid, content further include dilution, suspending liquid, antioxidant One or more of.
The dilution is selected from one or more of polyethylene glycol, propylene glycol, single diglycerine fatty acid ester, vegetable oil, Wherein vegetable oil may include one or more of compoundings such as peanut oil, soybean oil, rapeseed oil；It is preferred that polyethylene glycol and propylene glycol. It is more preferable using the compatibility of polyethylene glycol and propylene glycol as dilution and PUFA, and being capable of suspending SA particle to a certain extent.
It is sweet that the suspending liquid is selected from rosin glycerides, phosphatide, medium chain triglyceride, beeswax, sapn, tween, monostearate One or more compoundings in grease, bi-tristearin；It is preferred that one of medium chain triglyceride, beeswax, sapn and phosphatide Or two kinds.These types is good in oil compatibility, can not only play stabilization and can also adjust density of matrix and reach and preferably hold Carry effect.The suspending liquid is added directly into dispersing and dissolving in oil matrix.
The antioxidant is selected from ascorbic acid and its salt, arabo-ascorbic acid and its salt, ascorbic acid palm ester, vitamin E, one or more of 2,6 di tert butyl 4 methyl phenol (BHT), butylated hydroxy anisole (BHA)；
In the PUFA soft capsule, using polyethylene glycol as dilution, phosphatide is suspending liquid, ascorbyl palmitate When as antioxidant, the synergistic effect of above-mentioned auxiliary agent can be utilized, the stability and inoxidizability of whole system are obviously improved.
As the preferred technical solution of the present invention, a kind of PUFA soft capsule containing sialic acid is provided, in parts by weight, It is grouped as by following group:
80-100 parts of the PUFA oil suspension of the evenly dispersed sialic acid, 0-25 parts of dilution, 1-15 parts of suspending liquid, antioxygen 0-5 parts of agent；
It is highly preferred that
80-90 parts of the PUFA oil suspension of the evenly dispersed sialic acid, 3-15 parts of dilution, 1-15 parts of suspending liquid, antioxygen 0-0.8 parts of agent.
It is further preferred that the dilution is the 3.3-18.7wt% of the PUFA oil suspension；
And/or the suspending liquid is the 1.1-18.75wt% of the PUFA oil suspension；
And/or when the antioxidant PUFA oil suspension 0-1wt%.
Under normal circumstances, the additive amount of antioxidant is but provided by the present invention 1% or so in common PUFA grease In formula, since the overall coordination of active constituent, diluent and suspending liquid acts on, antioxidant can not be added, or few Amount addition.
In PUFA soft capsule of the present invention containing sialic acid, the active constituent of the PUFA oil suspension includes quality Than the sialic acid and PUFA for 1:(1.5-20) (mass ratio is with 1: (4-8) is more excellent)；
The active constituent can also further comprise carotenoid；The content of the carotenoid is no more than described The 20% of PUFA oil parts by weight；The more preferable carotenoid is bata-carotene, and bata-carotene is good compatible with oily phase Property and antioxygenic property, enable to system more stable.
As the highly preferred technical solution of the present invention, the PUFA soft capsule containing sialic acid, content include (by Following group is grouped as):
80-85 parts of PUFA oil suspension；4.5-15 parts of polyethylene glycol；0-2 parts of sapn, medium-chain fatty glyceride (MCT) 2- 15 parts.
It include sialic acid, DHA oil, ARA oil, carotenoid in the PUFA oil suspension.
The present invention provides the preparation method of soft capsule described in above-mentioned any one technical solution together, and steps are as follows:
Through pelleting by the content and utricule to obtain the final product.
The utricule is prepared or commercially available using the acceptable preparation method in this field；
There is provided a kind of more preferred preparation method herein: the utricule is former using gelatin, glycerol and pure water as utricule What material was prepared；
Specifically, by glycerol and pure water injection glue pot, it is heated with stirring to 60-80 DEG C, adds gelatin, continues to stir Vacuum demoulds after 20-30 minutes, and utricule is prepared.
It will be understood by those skilled in the art that above-mentioned preparation process may also include this field and prepare capsule in addition to pelleting Technological means, such as be formed, wash ball, drying, pick ball and packing step.
Soft capsule granule content uniform level provided by the present invention is high, and stability is high, portable and can supplement simultaneously more Kind nutrition, antioxidant addition is few or does not add, highly-safe, is suitable for pregnant baby crowd.Contain described in used in the present invention There is the PUFA oil suspension of sialic acid to be conducive to the nutritional supplementation substance of pregnant baby, and prepare easy, material and be easy to get, moderate cost produces Product are portable, are easy to save, and are suitable for promoting the use of.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..
Embodiment 1
The present embodiment provides a kind of PUFA oil suspension and preparation method thereof containing sialic acid, the PUFA oil are DHA oil； Specific step is as follows:
1) sialic acid powder 20g is taken, is mixed with the DHA oil of 160g, is sheared under the conditions of 17000rpm to abundant point It dissipates；
The different parts of oil suspension are taken to be measured after dispersion, the difference of sialic acid content is less than 3%；
2) sialic acid and DHA oil mixture after taking step 1) to disperse carry out high speed grinding, under nitrogen filled protection, with The speed of 2000r/min, sieve gap 0.4mm, the grinding 30mins time to obtain the final product.
Sialic acid powder described in the present embodiment, DHA oil derive from commercially available；Sialic acid content in the sialic acid powder Content >=35% of DHA during > 98%, the DHA are oily.
Embodiment 2
The present embodiment provides a kind of PUFA oil suspension and preparation method thereof containing sialic acid, the PUFA oil are DHA oil； Difference with embodiment 1 is only that:
Sialic acid powder employed in the present embodiment is prepared by the following method:
Using Escherichia coli as fermentation thalli, seed liquor is pressed 1% inoculum concentration by deposit number CCTCC NO:M 201810, It is added in the fermentor containing sterilization fermentation culture medium, LB culture medium contains: peptone 10g/L, yeast extract powder 5g/L, chlorine Change sodium 10g/L, pH7.0, ferment under conditions of 37 DEG C, by way of flow feeding, is carried out with the rate of 2.5g/ (L*h) Glucose is supplemented, puts tank after the 75h that continuously ferments, obtains the fermentation liquid containing sialic acid, fermented feed liquid divides in such a way that film filters Bacteria residue is separated out, the liquid being obtained by filtration is obtained using after sterilizing, decoloration, filtering, concentration, crystallization, separation, spray drying, sieving To sialic acid powder.
Sterilizing, decoloration, filtering, concentration, crystallization, separation, spray drying in above step and etc. according to this field Usual manner operation.
DHA oil described in the present embodiment is from commercially available；Content >=35% of DHA in the DHA oil.
Embodiment 3
The present embodiment provides a kind of PUFA oil suspension and preparation method thereof containing sialic acid, the PUFA oil are DHA oil； Difference with embodiment 2 is only that:
DHA oil employed in the present embodiment is prepared by the following method:
Using schizochytrium as fermentation thalli, the fermentor of the fermentation medium after seed liquor is added to containing steam sterilizing In, the formula of fermentation medium is as follows: glucose 100g/L, yeast extract 6g/L, sodium glutamate 20g/L, sodium chloride 3g/L, phosphorus Acid dihydride potassium 2g/L, magnesium sulfate 5g/L, calcium chloride 1g/L, sodium bicarbonate 0.5g/L, sodium sulphate 7g/L, ammonium sulfate 8g/L, chlorination Potassium 0.6g/L, pH are natural；The ratio 5% and dissolved oxygen ratio that seed liquor and fermentation medium are controlled in fermentation process are 8%, 28 Tank is put after DEG C fermentation 100h, and thallus is collected in centrifugation or filtering, adds 0.1% cellulase and 0.2% compound protease carries out Breaking-wall cell is centrifugally separating to obtain DHA crude oil, obtains two dodecahexaenes using degumming, alkali refining, decoloration, precipitation, deodorization etc. The grease of the DHA of 40% or more acid content.
Degumming described in above step, alkali refining, decoloration, precipitation, deodorization and etc. be all made of this field routine fermentation, Lock out operation.
Embodiment 4
The present embodiment provides a kind of PUFA oil suspension and preparation method thereof containing sialic acid, the PUFA oil are DHA oil； Specific step is as follows:
1) sialic acid is prepared:
Using Escherichia coli as fermentation thalli, seed liquor is pressed 1% inoculum concentration by deposit number CCTCC NO:M 201810, It is added in the fermentor containing sterilization fermentation culture medium, LB culture medium contains: peptone 10g/L, yeast extract powder 5g/L, chlorine Change sodium 10g/L, pH7.0, ferment under conditions of 37 DEG C, by way of flow feeding, is carried out with the rate of 2.5g/ (L*h) Glucose is supplemented, puts tank after the 75h that continuously ferments, obtains the fermentation liquid containing sialic acid, fermented feed liquid divides in such a way that film filters Bacteria residue is separated out, the liquid being obtained by filtration is obtained using after sterilizing, decoloration, filtering, concentration, crystallization, separation, spray drying, sieving To sialic acid powder.
Sterilizing, decoloration, filtering, concentration, crystallization, separation, spray drying in above step and etc. according to this field Usual manner operation.
2) preparation DHA oil:
Using schizochytrium as fermentation thalli, the fermentor of the fermentation medium after seed liquor is added to containing steam sterilizing In, the formula of fermentation medium is as follows: glucose 100g/L, yeast extract 6g/L, sodium glutamate 20g/L, sodium chloride 3g/L, phosphorus Acid dihydride potassium 2g/L, magnesium sulfate 5g/L, calcium chloride 1g/L, sodium bicarbonate 0.5g/L, sodium sulphate 7g/L, ammonium sulfate 8g/L, chlorination Potassium 0.6g/L, pH are natural；The ratio 5% and dissolved oxygen ratio that seed liquor and fermentation medium are controlled in fermentation process are 8%, 28 Tank is put after DEG C fermentation 100h, and thallus is collected in centrifugation or filtering, adds 0.1% cellulase and 0.2% compound protease carries out Breaking-wall cell is centrifugally separating to obtain DHA crude oil, obtains two dodecahexaenes using degumming, alkali refining, decoloration, precipitation, deodorization etc. The grease of the DHA of 40% or more acid content.
3) the DHA oil of the sialic acid powder 20g for taking step 1) to be prepared, the 100g being prepared with step 2) mix, It is sheared under the conditions of 17000rpm to fully dispersed；
4) sialic acid and DHA oil mixture after taking step 3) to disperse carry out high speed grinding, under nitrogen filled protection, with The speed of 2000r/min, sieve gap 0.4mm, the grinding 30mins time to obtain the final product.
Degumming described in above step, alkali refining, decoloration, precipitation, deodorization and etc. be all made of this field routine fermentation, Lock out operation.
Embodiment 5
The present embodiment provides a kind of PUFA oil suspension and preparation method thereof containing sialic acid, the PUFA oil are DHA oil； Specific step is as follows:
1) sialic acid is prepared:
Using Escherichia coli as fermentation thalli, seed liquor is pressed 1% inoculum concentration by deposit number CCTCC NO:M 201810, It is added in the fermentor containing sterilization fermentation culture medium, LB culture medium contains: peptone 10g/L, yeast extract powder 5g/L, chlorine Change sodium 10g/L, pH7.0, ferment under conditions of 37 DEG C, by way of flow feeding, is carried out with the rate of 2.5g/ (L*h) Glucose is supplemented, puts tank after the 75h that continuously ferments, obtains the fermentation liquid containing sialic acid, fermented feed liquid divides in such a way that film filters Bacteria residue is separated out, the liquid being obtained by filtration is obtained using after sterilizing, decoloration, filtering, concentration, crystallization, separation, spray drying, sieving To sialic acid powder.
Sterilizing, decoloration, filtering, concentration, crystallization, separation, spray drying in above step and etc. according to this field Usual manner operation.
2) preparation DHA oil:
Using schizochytrium as fermentation thalli, the fermentor of the fermentation medium after seed liquor is added to containing steam sterilizing In, glucose 100g/L, yeast extract 6g/L, sodium glutamate 20g/L, sodium chloride 3g/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 5g/L, calcium chloride 1g/L, sodium bicarbonate 0.5g/L, sodium sulphate 7g/L, ammonium sulfate 8g/L, potassium chloride 0.6g/L, pH are natural；Fermentation The ratio 5% and dissolved oxygen ratio for controlling seed liquor and fermentation medium in the process are 8%, put tank after 28 DEG C of fermentation 100h, from Thallus is collected in the heart or filtering, adds 0.1% cellulase and 0.2% compound protease carries out breaking-wall cell, is centrifugated To DHA crude oil, the DHA of 40% or more docosahexaenoic acid content is obtained using degumming, alkali refining, decoloration, precipitation, deodorization etc. Grease.
3) the DHA oil of the sialic acid powder 40g for taking step 1) to be prepared, the 160g being prepared with step 2) mix, It is sheared under the conditions of 18000rpm to fully dispersed；
4) sialic acid and DHA oil mixture after taking step 3) to disperse carry out high speed grinding, under nitrogen filled protection, with The speed of 2500r/min, sieve gap 0.35mm, the grinding 25mins time to obtain the final product.
Degumming described in above step, alkali refining, decoloration, precipitation, deodorization and etc. be all made of this field routine fermentation, Lock out operation.
Embodiment 6
The present embodiment provides a kind of PUFA oil suspension and preparation method thereof containing sialic acid, the PUFA oil are DHA oil； Specific step is as follows:
1) sialic acid is prepared:
Using Escherichia coli as fermentation thalli, seed liquor is pressed 1% inoculum concentration by deposit number CCTCC NO:M 201810, It is added in the fermentor containing sterilization fermentation culture medium, LB culture medium contains: peptone 10g/L, yeast extract powder 5g/L, chlorine Change sodium 10g/L, pH7.0, ferment under conditions of 37 DEG C, by way of flow feeding, is carried out with the rate of 2.5g/ (L*h) Glucose is supplemented, puts tank after the 75h that continuously ferments, obtains the fermentation liquid containing sialic acid, fermented feed liquid divides in such a way that film filters Bacteria residue is separated out, the liquid being obtained by filtration is obtained using after sterilizing, decoloration, filtering, concentration, crystallization, separation, spray drying, sieving To sialic acid powder.
2) preparation DHA oil:
Using schizochytrium as fermentation thalli, the fermentor of the fermentation medium after seed liquor is added to containing steam sterilizing In, glucose 100g/L, yeast extract 6g/L, sodium glutamate 20g/L, sodium chloride 3g/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 5g/L, calcium chloride 1g/L, sodium bicarbonate 0.5g/L, sodium sulphate 7g/L, ammonium sulfate 8g/L, potassium chloride 0.6g/L, pH are natural；Fermentation The ratio 5% and dissolved oxygen ratio for controlling seed liquor and fermentation medium in the process are 8%, put tank after 28 DEG C of fermentation 100h, from Thallus is collected in the heart or filtering, adds 0.1% cellulase and 0.2% compound protease carries out breaking-wall cell, is centrifugated To DHA crude oil, the DHA of 40% or more docosahexaenoic acid content is obtained using degumming, alkali refining, decoloration, precipitation, deodorization etc. Grease.
3) the DHA oil of the sialic acid powder 40g for taking step 1) to be prepared, the 80g being prepared with step 2) mix, It is sheared under the conditions of 18000rpm to fully dispersed；
4) sialic acid and DHA oil mixture after taking step 3) to disperse carry out high speed grinding, under nitrogen filled protection, with The speed of 2500r/min, sieve gap 0.35mm, the grinding 30mins time to obtain the final product.
Embodiment 7
The present embodiment provides a kind of PUFA oil suspension and preparation method thereof containing sialic acid, the PUFA oil are DHA oil With the miscella of ARA oil；Specific step is as follows:
1) sialic acid is prepared:
Using Escherichia coli as fermentation thalli, seed liquor is pressed 1% inoculum concentration by deposit number CCTCC NO:M 201810, It is added in the fermentor containing sterilization fermentation culture medium, LB culture medium contains: peptone 10g/L, yeast extract powder 5g/L, chlorine Change sodium 10g/L, pH7.0, ferment under conditions of 37 DEG C, by way of flow feeding, is carried out with the rate of 2.5g/ (L*h) Glucose is supplemented, puts tank after the 75h that continuously ferments, obtains the fermentation liquid containing sialic acid, fermented feed liquid divides in such a way that film filters Bacteria residue is separated out, the liquid being obtained by filtration is obtained using after sterilizing, decoloration, filtering, concentration, crystallization, separation, spray drying, sieving To sialic acid powder.
2) preparation DHA oil:
Using schizochytrium as fermentation thalli, the fermentor of the fermentation medium after seed liquor is added to containing steam sterilizing In, the formula of the fermentation medium is as follows: glucose 100g/L, yeast extract 6g/L, sodium glutamate 20g/L, sodium chloride 3g/ L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 5g/L, calcium chloride 1g/L, sodium bicarbonate 0.5g/L, sodium sulphate 7g/L, ammonium sulfate 8g/L, Potassium chloride 0.6g/L, pH are natural；Seed liquor is controlled in fermentation process and the ratio 5% and dissolved oxygen ratio of fermentation medium are Tank, centrifugation or filtering are put after 8%, 28 DEG C of fermentation 100h, collects thallus, adjust pH value to 11.5,0.1%g basic protein is added Enzyme, 10% phosphatide, digest 5h by 40 DEG C of temperature control, and film is added in the mixture after the mixture, cell cracking after obtaining cell cracking Filter plant, circulating filtration isolate the emulsified fat containing moisture；
The sodium chloride of 3.0% missible oil weight is added into the emulsified fat containing moisture, sedimentation takes supernatant after stirring and evenly mixing Liquid obtains the grease of the DHA of 40% or more docosahexaenoic acid content.3) preparation ARA oil
After the bacterium in 50m3 seeding tank is dense reach 30% (volume ratio) after, be linked by culture transferring pipeline equipped with 130m3Hair The 200m of ferment culture medium3It is cultivated in fermentor, inoculum concentration 20% (volume ratio), fermentor controls 30 DEG C of temperature, stirring speed 300 revs/min, ventilatory capacity 1.5vvm (L/L.min) of degree, tank press 0.15Mpa, cultivate 180h.Pass through stream plus carbon in fermentation process Source controls in fermentation liquid carbon source concentration in 20g/L, the fermentation tank culture medium are as follows: carbon source starch 50g/l；Nitrogen source yeast extract 20g/l；pH 8.5.The separation of fermentative broth that fermented and cultured obtains, obtains wet thallus, and drying obtains dry mycelium 50g.Into dry mycelium Extractant n-hexane is added to be extracted, isolated solid formation, which is transferred in extraction container, after extraction carries out repeating extraction, such as This, terminates extraction process, 200 milliliters of n-hexanes is added when extracting for the first time, be added every time later when until oil-free in extract liquor 150 milliliters of n-hexanes, the miscella being separated by filtration after each extraction is filled, precipitation obtain ARA grease.
4) the DHA oil and step of the sialic acid powder 30g, the 160g being prepared with step 2) that take step 1) to be prepared 3) the 80gARA mixing being made, is sheared under the conditions of 18000rpm to fully dispersed；
5) sialic acid and DHA/ARA oil mixture after taking step 4) to disperse carry out high speed grinding, under nitrogen filled protection, with The speed of 2500r/min, sieve gap 0.35mm, the grinding 30mins time to obtain the final product.
Embodiment 8
The source of SA and DHA and preparation method be with embodiment 3 in the present embodiment oil suspension, the difference is that, in step 3 Before added with 10% beta carotene for accounting for the PUFA oil gross mass, added during step 3.
Embodiment 9
The source of SA, DHA and ARA and preparation method be with embodiment 7 in the present embodiment oil suspension, the difference is that, It is added with 1.7% beta carotene before step 3, is added during step 3, and sialic acid powder is 20g, DHA+ ARA oil is 80g.
After oil suspension obtained by above-described embodiment 1-9 is prepared is placed for 24 hours, through the sialic acid content for measuring variant position Difference is below 5%.
Embodiment 10
The present embodiment provides a kind of PUFA soft capsule containing sialic acid, content include:
Embodiment 11
The present embodiment provides a kind of PUFA soft capsule containing sialic acid, content include:
Embodiment 12
The present embodiment provides a kind of PUFA soft capsule containing sialic acid, content include:
Embodiment 13
The present embodiment provides a kind of PUFA soft capsule containing sialic acid, content include:
Embodiment 14
The present embodiment provides a kind of PUFA soft capsule containing sialic acid, content include:
Embodiment 15
The present embodiment provides a kind of PUFA soft capsule containing sialic acid, content include:
PUFA oil suspension provides 85 parts by embodiment 9；
15 parts of dilution propylene glycol.
Embodiment 16
The present embodiment provides a kind of PUFA soft capsules and preparation method thereof, specifically, include the following steps:
1. 10 parts of 82.5 parts of the PUFA oil suspension and dilution polyethylene glycol of the evenly dispersed sialic acid of Example 3, 5 parts of 2 parts+medium-chain fatty glyceride of suspending liquid sapn, E0.5 parts of antioxidant vitamin be content；
2. preparing utricule: using gelatin, glycerol and pure water as utricule raw material, by glycerol and pure water injection glue pot, stirring It mixes and is heated to 60-80 DEG C, add gelatin, vacuum demoulds after continuing stirring 20-30 minutes, and utricule is prepared；
3. preparing capsule: by content prepared by step 1 and utricule prepared by step 2 by pelleting, be formed, wash ball, dry It is dry, pick ball and packing step, PUFA soft capsule is prepared.
Embodiment 17
The present embodiment provides a kind of PUFA soft capsules and preparation method thereof, specifically, include the following steps:
1. preparing content: evenly dispersed 84.5 parts of the PUFA oil suspension and polyethylene glycol of sialic acid of Example 7 10 parts, 5 parts of medium chain triglyceride, 0.5 part of vitamin E be content；
2. preparing utricule: using 10 parts of gelatin, 3 parts of glycerol and 10 parts of pure water as utricule raw material, glycerol and pure water being injected In glue pot, it is heated with stirring to 60-80 DEG C, adds gelatin, vacuum demoulds after continuing stirring 20-30 minutes, and capsule is prepared Body；
3. preparing capsule: by content prepared by step 1 and utricule prepared by step 2 by pelleting, be formed, wash ball, dry It is dry, pick ball and packing step, PUFA soft capsule is prepared.
Embodiment 18
The present embodiment provides a kind of PUFA soft capsules and preparation method thereof, specifically, include the following steps:
1. preparing content: evenly dispersed 85 parts of the PUFA oil suspension and polyethylene glycol 8 of sialic acid of Example 7 Part, 5 parts of 2 parts+medium-chain fatty glyceride of sapn (MCT) be content；
2. preparing utricule: using 10 parts of gelatin, 3 parts of glycerol and 10 parts of pure water as utricule raw material, glycerol and pure water being injected In glue pot, it is heated with stirring to 60-80 DEG C, adds gelatin, vacuum demoulds after continuing stirring 20-30 minutes, and capsule is prepared Body；
3. preparing capsule: by content prepared by step 1 and utricule prepared by step 2 by pelleting, be formed, wash ball, dry It is dry, pick ball and packing step, PUFA soft capsule is prepared.
Embodiment 19
The present embodiment provides PUFA soft capsules and preparation method thereof described in embodiment 14 specifically to include the following steps:
1. preparing content: evenly dispersed 85 parts of the PUFA oil suspension and polyethylene glycol 4.5 of sialic acid of Example 9 Part, 10 parts of medium-chain fatty glyceride (MCT), 0.5 part of ascorbyl palmitate be content；
2. preparing utricule: using 10 parts of gelatin, 3 parts of glycerol and 10 parts of pure water as utricule raw material, glycerol and pure water being injected In glue pot, it is heated with stirring to 60-80 DEG C, adds gelatin, vacuum demoulds after continuing stirring 20-30 minutes, and capsule is prepared Body；
3. preparing capsule: by content prepared by step 1 and utricule prepared by step 2 by pelleting, be formed, wash ball, dry It is dry, pick ball and packing step, PUFA soft capsule is prepared.
Comparative example 1
This comparative example provides a kind of DHA oil suspension and preparation method thereof containing sialic acid, compared with Example 1 compared with area It is not only that, sialic acid powder usage amount is 80g, and DHA oil usage amount is 40g.
Comparative example 2
This comparative example provides a kind of DHA oil suspension and preparation method thereof containing sialic acid, compared with Example 1 compared with area It is not only that, step 2) is replaced with is dispersed using ultrasonic power, ultrasound condition 25kHz, is carried out 20 minutes.
Comparative example 3
This comparative example provides a kind of PUFA soft capsule containing sialic acid, and content includes:
Comparative example 4
This comparative example provides a kind of PUFA soft capsule containing sialic acid, and content includes:
Comparative example 5
This comparative example provides a kind of PUFA soft capsule containing sialic acid, and content includes:
Comparative example 6
This comparative example provides a kind of PUFA soft capsule, and content includes:
Test example 1
This test example provides the stability test of soft capsule provided by embodiment 8-13, includes hot test, Qiang Guangshi It tests and long term test.
1, capsule quality is examined
1.1 are detected 9-14 of the embodiment of the present invention and comparative example 3-5 soft capsule prepared by emphasis quality index, point Its quality change situation is not investigated, and content and related substance are detected with methanol: water (3: 1) is mobile phase, and Detection wavelength is 240nm is checked with efficient form and aspect chromatography.
1.2 the results are shown in Table 1.
1 capsule quality of table detects (every 100)
2, long-term stable experiment
2.1 soft capsules for preparing 9-14 of the embodiment of the present invention and comparative example 3-6 are at 25 DEG C ± 2 DEG C of temperature, relative humidity It places under the conditions of 60% ± 10%, respectively at the 3rd, inspection by sampling in 6 months, is detected by emphasis quality index for a long time, respectively Its quality change situation is investigated,
2.2 the results are shown in Table 2.
2 long-term stable experiment result of table
Table 2 the results show that the soft capsule sample of 8-14 of embodiment of the present invention preparation through 25 DEG C ± 2 DEG C of temperature, relative humidity It is placed for a long time 6 months under the conditions of 60% ± 10%, compared with 0 day, every quality index variation is less, relatively stable.
Although above having used general explanation, specific embodiment and test, the present invention is made to retouch in detail It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art 's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed Range.

Claims (10)

1. a kind of PUFA soft capsule containing sialic acid, which is characterized in that content includes: the PUFA oil of evenly dispersed sialic acid Suspension.
2. soft capsule according to claim 1, which is characterized in that the PUFA oil suspension is that sialic acid is added to PUFA oil In be dispersed up to take oil suspension different parts measurement sialic acid content, content difference control within 5% to obtain the final product；
Preferably, the PUFA oil suspension of the evenly dispersed sialic acid is prepared by following methods:
The dispersing mode, which uses, first to be sheared, grinds again；
When shearing, 10-35min is carried out under the shear velocity that condition is 10000rpm-20000rpm, it is highly preferred that condition is 10-20mins is carried out under the shear velocity of 15000rpm-18000rpm；
Condition when grinding are as follows: circular grinding 10-50min under revolution 1000r/min-4000r/min, 0.2-0.9mm strainer； It is highly preferred that being 2000r/min-3000r/min, the circular grinding 20-30mins under 0.4-0.6mm strainer in revolution.
3. PUFA soft capsule according to claim 1 or 2, which is characterized in that the content further includes dilution, suspending One or more of liquid, antioxidant；
Preferably, the PUFA soft capsule is in parts by weight, comprising:
80-100 parts of the PUFA oil suspension of the evenly dispersed sialic acid, 0-25 parts of dilution, 1-15 parts of suspending liquid, antioxidant 0-5 parts；
It is highly preferred that
80-90 parts of the PUFA oil suspension of the evenly dispersed sialic acid, 3-15 parts of dilution, 1-15 parts of suspending liquid, antioxidant 0-0.8 parts.
4. PUFA soft capsule according to claim 3, which is characterized in that in the content, the dilution is selected from poly- One or more of ethylene glycol, propylene glycol, single diglycerine fatty acid ester, vegetable oil；It is preferred that polyethylene glycol and/or propylene glycol；
And/or the suspending liquid is selected from rosin glycerides, phosphatide, medium chain triglyceride, beeswax, sapn, tween, monostearate One or more compoundings in glyceride, bi-tristearin；It is preferred that one in medium chain triglyceride, beeswax, sapn and phosphatide Kind or two kinds；
And/or the antioxidant is selected from ascorbic acid and its salt, arabo-ascorbic acid and its salt, ascorbic acid palm ester, dimension life One or more of plain E, 2,6 di tert butyl 4 methyl phenol, butylated hydroxy anisole.
5. PUFA soft capsule according to claim 3 or 4, which is characterized in that the dilution is the PUFA oil suspension 3.3-18.7wt%；
And/or the suspending liquid is the 1.1-18.75wt% of the PUFA oil suspension；
And/or when the antioxidant PUFA oil suspension 0-1wt%.
6. PUFA soft capsule according to claim 1-5, which is characterized in that the evenly dispersed sialic acid PUFA oil suspension includes that mass ratio is oily for the sialic acid and PUFA of 1:(1.5-20)；It is preferred that 1: (4-8).
7. PUFA soft capsule according to claim 1-6, which is characterized in that the evenly dispersed sialic acid In PUFA oil suspension, the PUFA oil includes at least one of DHA, ARA, linolenic acid, linoleic acid；
Preferably, the PUFA oil includes at least one of DHA oil or ARA oil；
It is highly preferred that the content ratio of the DHA oil and ARA oil is (2- in the PUFA oil suspension of the evenly dispersed sialic acid 3): 1, and the total content of DHA oil and ARA oil accounts at least the 80% of the PUFA oil.
8. PUFA soft capsule according to claim 7, which is characterized in that the PUFA oil suspension of the evenly dispersed sialic acid In further include carotenoid；
Preferably, the content of the carotenoid is no more than the 20% of the PUFA oil parts by weight.
9. PUFA soft capsule according to claim 1-8, which is characterized in that in parts by weight, comprising: PUFA 80-85 parts of oil suspension；4.5-15 parts of polyethylene glycol；0-2 parts of sapn, 2-15 parts of medium-chain fatty glyceride；
It include sialic acid, DHA oil, ARA oil, carotenoid in the PUFA oil suspension.
10. a kind of method for preparing the described in any item PUFA soft capsules of claim 1-9, which is characterized in that by the content Object and utricule through pelleting to obtain the final product；
Preferably, the utricule is prepared using gelatin, glycerol and pure water as utricule raw material；Glycerol and pure water are injected In glue pot, it is heated with stirring to 60-80 DEG C, adds gelatin, vacuum demoulds after continuing stirring 20-30 minutes, and capsule is prepared Body.
CN201811358736.0A 2018-11-15 2018-11-15 A kind of PUFA soft capsule and preparation method thereof containing sialic acid Pending CN109601980A (en)

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CN112493477A (en) * 2020-11-30 2021-03-16 嘉必优生物技术（武汉）股份有限公司 Use of a sialylated oil suspension for the preparation of capsules
CN112535292A (en) * 2020-11-30 2021-03-23 嘉必优生物技术（武汉）股份有限公司 Sialic acid oil suspension, and preparation method and application thereof
CN118147023A (en) * 2024-05-13 2024-06-07 山东润德生物科技有限公司 Composite starter and application thereof in preparation of N-acetylneuraminic acid

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CN112493477B (en) * 2020-11-30 2022-09-13 嘉必优生物技术（武汉）股份有限公司 Use of a sialylated oil suspension for the preparation of capsules
CN118147023A (en) * 2024-05-13 2024-06-07 山东润德生物科技有限公司 Composite starter and application thereof in preparation of N-acetylneuraminic acid
CN118147023B (en) * 2024-05-13 2024-07-19 山东润德生物科技有限公司 Composite starter and application thereof in preparation of N-acetylneuraminic acid

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Application publication date: 20190412